In silico Analysis, Molecular Docking, Molecular Dynamic, Cloning, Expression and Purification of Chimeric Protein in Colorectal Cancer Treatment

Drug Design, Development and Therapy
Hassan DanaVahid Marmari

Abstract

Colorectal cancer (CRC) is a type of cancer in humans that leads to high mortality and morbidity. CD166 and CD326 are immunoglobulins that are associated with cell migration. These molecules are included in tumorigenesis of CRC and serve a great marker of CRC stem cells. In the present study, we devised a novel chimeric protein including the V1-domain of the CD166 and two epitopes of CD326 to use in diagnostic or therapeutic applications. In silico techniques were launched to characterize the properties and structure of the protein. We have predicted physicochemical properties, structures, stability, MHC class I binding properties and ligand-receptor interaction of this chimeric protein by means of computational bioinformatics tools and servers. The sequence of chimeric gene was optimized for expression in prokaryotic host using online tools and cloned into pET-28a plasmid. The recombinant pET28a was transformed into the E. coli BL21DE3. Expression of recombinant protein was examined by SDS-PAGE and Western blotting. The designed chimeric protein retained high stability and the same immunogenicity as of the original proteins. Bioinformatics data indicated that the epitopes of the synthetic chimeric protein might induce B-cell- ...Continue Reading

Citations

Nov 24, 2020·Journal of Biomolecular Structure & Dynamics·Zahra YazdaniReza Valadan
Feb 26, 2021·Clinical and Experimental Vaccine Research·Asqwin Uthaya KumarAnna Pick Kiong Ling
Apr 9, 2021·Life Sciences·Pawan Kumar Raghav, Zoya Mann

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Methods Mentioned

BETA
in silico techniques
ELISA

Software Mentioned

Modeller9
ProtParam
Vaxijen
NetSurfP
SASA
ExPASy ProtParam
pBLAST
DSSP
SOPMA
Raptor X

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